Talk:DNA ligation

General ligation protocols are fine for protruding sticky end ligations but often fail with blunt ends or recessed sticky ends. For blunt ends, higher enzyme concentrations are recommended along with considerably lower total DNA concentrations (cf. Bercovich et al., BioTechniques 12(2), 190-193 (1992)). If there is someone around with a good working protocol for ligation of blunt and of protruding ends, it would be much appreciated. {MDolinar, July 25, 2007}

Insert to Vector ratios

Hi, new to this wiki but not to wiki's. I notice that a formula was used for a 6:1 insert:vector ratio at the top of the page but later it talks about the appropraite range is 1:1 to 5:1, this seems like a contradiction. Under what conditions does one use the 6:1 ratio. This page needs at least some discussion of how to ligate vectors to inserts where both are reasonably long. Phil 18:22, 6 September 2007 (EDT).

• Reshma 19:50, 6 September 2007 (EDT): Good catch. I think this may be a reflection of the fact that depending on exactly what you are ligating, the optimal insert:vector ratio can differ so different people advocate different ratio's. Hence the note in bold regarding the insert:vector ratio. If you have suggestions on ligating long inserts, please feel free to add it to the Notes section.